CN114107184A - 一种siRNA在牙髓干细胞成骨分化中的应用 - Google Patents
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Abstract
本发明公开了一种siRNA在牙髓干细胞成骨分化中的应用,属于干细胞技术领域。所述siRNA为LINC02192的siRNA,所述siRNA的序列为Sense:GGUGGAGGUUGCUAAGCAAAG;Antisense:UUGCUUAGCAACCUCCACCUG。使用本发明siRNA转染牙髓干细胞后,能够有效的促进牙髓干细胞的成骨分化相关蛋白的表达并能够有效的促进牙髓干细胞的成骨分化。
Description
技术领域
本发明属于干细胞技术领域,尤其涉及一种用于牙髓干细胞成骨分化和增殖的siRNA。
背景技术
牙髓组织位于牙齿内部的牙髓腔内,是牙体组织中唯一的软组织。2000年Gronthos 等通过对人牙髓细胞的研究,发现了一种与骨髓间充质干细胞有着极其相似的免疫表型及形成矿化结节能力的细胞,细胞中形态呈梭形,可自我更新和多向分化,有着较强的克隆能力。这些由牙髓组织中分离出的成纤维状细胞就称为牙髓干细胞 (DentalPulp Stem Cells, DPSCs)。
牙髓干细胞具有多向分化的潜能,可以分化为各种细胞,如成骨细胞、成牙本质细胞、成软骨细胞、成脂肪细胞、神经细胞和成肌细胞。其中,将牙髓干细胞进行成骨分化诱导来修复牙齿,是牙髓干细胞的主要应用方向,因此实现牙髓干细胞的快速成骨诱导有助于牙髓干细胞的大规模医学应用。
发明内容
本发明的目的在于提供一种用于促进牙髓干细胞成骨分化的siRNA。
为实现上述目的,本发明提供了如下技术方案:
其一,本发明提供了一种siRNA在制备牙髓干细胞成骨分化促进剂中的用途,所述siRNA为LINC02192的siRNA。
优选地,所述LINC02192的序列如SEQ ID NO.1所示。
优选地,所述siRNA的序列如SEQ ID NO.2和SEQ ID NO.3所示。
其二,本发明提供了一种用于促进牙髓干细胞成骨分化的siRNA,所述siRNA为LINC02192的siRNA。
优选地,所述LINC02192的序列如SEQ ID NO.1所示;所述siRNA的序列如SEQ IDNO.2和SEQ ID NO.3所示。
其三,本发明提供了一种siRNA在制备牙髓干细胞成骨分化相关蛋白ALP,RUNX2和OPN蛋白表达促进剂中的用途,所述siRNA为LINC02192的siRNA。
优选地,所述LINC02192的序列如SEQ ID NO.1所示;所述siRNA的序列如SEQ IDNO.2和SEQ ID NO.3所示。
其四,本发明提供了一种促进牙髓干细胞成骨分化的方法,其特征在于,包括如下步骤:
(1)将原代培养或传代培养获得的牙髓干细胞接种于培养装置中;
(2)将LINC02192的siRNA导入到牙髓干细胞中,加入成骨诱导培养基进行培养。
优选地,所述siRNA的序列如SEQ ID NO.2和SEQ ID NO.3所示。
本发明的有益效果是:
本发明的有益效果在于本发明发现LINC02192的siRNA能够有效的促进牙髓干细胞成骨分化相关蛋白的表达,并能够有效的促进牙髓干细胞成骨分化,因此,可将其用于牙髓干细胞成骨分化促进剂。
附图说明
图1 si-LINC02192的抑制效率检测结果;
图2 转染si-LINC02192后,成骨分化蛋白ALP,RUNX2和OPN的表达变化情况;
图3 转染si-LINC02192后,细胞钙化变化情况;
图4 转染si- LINC02192后,ALP染色变化情况。
具体实施方式
为能清楚说明本方案的技术特点,下面通过具体实施方式,对本方案进行阐述。
实施例1 LINC02192的siRNA对于成骨分化相关蛋白ALP,RUNX2和OPN的影响
1.设计合成LINC02192的siRNA (si-LINC02192)
LINC02192(序列如SEQ ID NO.1所示)的siRNA的序列如下所示:
Sense:GGUGGAGGUUGCUAAGCAAAG,SEQ ID NO.2
Antisense:UUGCUUAGCAACCUCCACCUG, SEQ ID NO.3
2.验证si-LINC02192的抑制效果
(1)将生长状态良好的1.5×104的第三代人牙髓干细胞接种于6孔板中;
(2)待细胞密度达到80%时,将si-NC和si-LINC02192的脂质体复合物加入到6孔板中;
(3)6h后,弃去上清,加入2ml完全培养基;
(4)培养48h后,弃去培养基,使用PBS洗涤细胞3次,每孔加入1mlTrizol试剂,反复吹打细胞将细胞吹下后,室温静置5min;
(5)将混合液转移至1.5ml的EP管中,加入200μl的氯仿,盖紧EP管后,手动剧烈震荡15s,室温放置5min;
(6)调整低温离心机的参数为4℃,12000g,15min,进行离心,离心后液体分为3层,小心的吸取上层的水相至新的EP管中;
(7)在新的EP管中加入0.5ml的异丙醇,混匀后静置10min,调整低温离心机的参数为4℃,12000g,10min,进行离心;
(8)弃去上清液,在沉淀中加入75%乙醇洗涤沉淀,调整离心机参数为4℃,7500g,5min,进行离心;
(9)弃去上清,置于超净台中空气干燥5-10min,至酒精完全挥发,加入40μl无RNAase的水,溶解沉淀,-80℃冰箱保存。
(10)去除基因组DNA
配置的反应体系如下:
反应体系 | 添加量 |
5×gDNA Eraser Buffer | 2μl |
gDNA Eraser | 1μl |
Total RNA | 1μg |
RNase Free dH2O | up to 10μl |
设定的反应条件如下:42℃ 2min,4℃ forever;
(11)逆转录反应
配置的反应体系如下:
反应体系 | 添加量 |
步骤(10)所得反应液 | 10μl |
PrimeScript RT Enzyme Mix I | 1μl |
5×PrimeScript Buffer 2 | 4μl |
RT Primer Mix | 1μl |
RNase Free dH2O | 4μl |
(12)荧光定量RCR反应
反应体系 | 添加量 |
SYBR Green Premix Ex Taq(2×) | 10μl |
正向引物 | 0.4μl |
反向引物 | 0.4μl |
cDNA模板 | 2μl |
ddH2O | 7.2μl |
设定的反应条件为:95℃ 5min;95℃ 10s,60℃ 30s,72℃ 35s,35个循环;
引物序列如下:
数据采用2-△△ CT法进行分析。
结果如图1所示,可以看出转染si-LINC02192后的相对表达量为0.33±0.05,说明si-LINC02192能够有效的抑制LINC02192的表达水平,因此si-LINC02192对于抑制LINC02192的表达是有效的。
3. 成骨分化过程中,转染si-LINC02192对于成骨分化相关蛋白表达的影响
(1)将生长状态良好的1.5×104的第三代人牙髓干细胞接种于6孔板中;
(2)待细胞密度达到80%时,将si-NC和si-LINC02192的脂质体复合物加入到6孔板中;
(3)6h后,弃去上清,加入2ml成骨分化培养基培养7天;
(4)培养结束后,去除培养基加入100μl细胞裂解液,收集裂解液至EP管中,冰上裂解30min,12000rpm/min离心20min后取上清;
(5)BCA法测量蛋白浓度后,制备成蛋白样品;
(6)配置5%浓缩胶和12%分离胶,每孔加入20μg蛋白样品,恒压80V至分离胶,120V至溴酚蓝至分离胶底部;
(7)将PVDF膜置于甲醇中浸泡15s,活化后安装电转夹,恒流250mA 1.5h,将PVDF膜取出置于5%脱脂奶粉中,封闭1h;
(8)孵育一抗,置于4℃孵育过夜,孵育结束后,TBST清洗PVDF膜3次,每次10min;
(9)取出PVDF膜,孵育2抗,室温条件下孵育60min,孵育结束后,TBST清洗PVDF膜3次,每次10min;
(10)在避光条件下,对PVDF膜进行显像后保存图像。
结果如图2,从图中可以看出,使用siRNA抑制LINC02192后,成骨分化相关蛋白ALP,RUNX2和OPN的蛋白表达量显著增加,说明si-LINC02192对于牙髓干细胞成骨分化相关蛋白的表达具有促进作用。
实施例2 LINC02192的siRNA对于牙髓干细胞成骨分化具有促进作用
1.细胞矿化检测
(1)将生长状态良好的1.5×104的第三代人牙髓干细胞接种于6孔板中;
(2)待细胞密度达到80%时,将si-NC和si-LINC02192的脂质体复合物加入到6孔板中;
(3)6h后,弃去上清,加入2ml成骨分化培养基培养14天;
(4)培养14天后,去除培养基,使用PBS轻轻的清洗细胞,加入4%多聚甲醛固定细胞30min;
(5)弃去4%多聚甲醛,使用PBS清洗细胞,使用茜素红染液室温避光孵育30min;
(6)去除茜素红染色液,使用PBS清洗2次,进行拍照。
2.细胞ALP染色检测
(1)将生长状态良好的1.5×104的第三代人牙髓干细胞接种于6孔板中;
(2)待细胞密度达到80%时,将si-NC和si-LINC02192的脂质体复合物加入到6孔板中;
(3)6h后,弃去上清,加入2ml成骨分化培养基培养7天;
(4)培养7天后,去除培养基,使用PBS轻轻的清洗细胞,加入4%多聚甲醛固定细胞30min;
(5)弃去4%多聚甲醛,使用PBS清洗细胞,参照BCIP NBT碱性磷酸酶显色试剂盒说明书处理细胞;
(6)处理结束后,于显微镜下进行拍照。
细胞矿化结果如图3所示,从图中可以看出,使用siRNA抑制LINC02192后,牙髓干细胞矿化结节的数量显著增加,说明si-LINC02192对于牙髓干细胞细胞矿化具有促进作用;
细胞ALP染色结果如图4,从图中可以看出,使用siRNA抑制LINC02192后,牙髓干细胞ALP染色程度显著升高,说明si-LINC02192对于ALP酶的表达具有促进作用;
从上述结果可以看出,说明si-LINC02192对于牙髓干细胞成骨分化具有促进作用。
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<213> 人源(Human)
<400> 1
ttaacatcac ttccccaggg cgaggaactc catgtgaggc tctgggggaa cctgactatg 60
tgcagtcgtt tacagcctgg gtctcctgga gcctatgaaa tgagaagcag tcattagcac 120
agcaagcggc cctcatcctt ccttctcaat aataacactg tcaccagagg atggaaaatt 180
accctgacgt ctctgtatct ccaaattgca cttcagagac agactgcgta aggcaaacag 240
accctgagtg aaaacctcct ggctagactt ccaagccaag gcctaataaa taaccctgaa 300
atatacaaag tgatggcagc tttgctctgt ggctcgtggc ttctgccctg ggaaggacca 360
tgaatttcag cgtctgacag atggggtcag aaagtggagt ctgcctggtg ggttgaggat 420
ctgcaaatta accccatggt ggtggtttca gaacaagtgt gcctgctcac gcaaagacta 480
acctctaaga aaggagcagg cccggaggtg aggccccctg ggcgtgaaag aatcaacccc 540
agtccacaaa gtccggagat gagacatcct gcctggagga gggagccctg tgcatcgcag 600
gatcaaagaa gccaaacagg atctttagtg gtgaccgaaa atgctcatga ggagaaaggc 660
tgcatggcat taagtgaacc ccattgaaag ggcccagtga acagagcccc tggacattgc 720
cggaaggaaa ggagaaagcc cagcaaagca cacaacgtat caggcttttc atgtgtcatt 780
gggtgaaagg gagtcacatg ggccaaggag gggaagcagg tgtcatcaga gcagttccac 840
agccctctag gcacagtaac aggcatgctt tctgtccttc tctcctttta gattgtaagc 900
tacccaaagt ccatctccat gggttttttt ccttatgtgc aaactaccat atgacaggtg 960
tgcctgacaa taactcaggt atagctgaga atgatcctgt agtccaagaa tgttggttct 1020
gagctctgaa ctaaggaatc tgggagctgc caacccagag gtttactcct tatctatgga 1080
gcataggtga acccctggcc catttcttgg aacagcatgt gcggggaacc aaggcccttt 1140
gttttgagct aggtggaggt ggccaggtag aggtcgccag gaagaggtgg ccaggtggag 1200
gttgctaagc aaagattgct atattaactg ggtgcttttt agaaaccata gtggttaccc 1260
cattcatcca gccactcctg gactccctgt acgtttaagt tgcccaataa aacttatgtc 1320
ttgctaaaaa aaaaaaaaaa a 1341
<210> 2
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 2
gguggagguu gcuaagcaaa g 21
<210> 3
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 3
uugcuuagca accuccaccu g 21
<210> 4
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
cggagatgag acatcctgcc 20
<210> 5
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
ccatgcagcc tttctcctca 20
<210> 6
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
acccactcct ccacctttga 20
<210> 7
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
ctgttgctgt agccaaattc gt 22
Claims (9)
1.一种siRNA在制备牙髓干细胞成骨分化促进剂中的用途,其特征在于,所述siRNA为LINC02192的siRNA。
2.根据权利要求1所述的用途,其特征在于,所述LINC02192的序列如SEQ ID NO.1所示。
3.根据权利要求1所述的用途,其特征在于,所述siRNA的序列如SEQ ID NO.2和SEQ IDNO.3所示。
4.一种用于促进牙髓干细胞成骨分化的siRNA,其特征在于,所述siRNA为LINC02192的siRNA。
5.根据权利要求1所述的siRNA,其特征在于,所述LINC02192的序列如SEQ ID NO.1所示;所述siRNA的序列如SEQ ID NO.2和SEQ ID NO.3所示。
6.一种siRNA在制备牙髓干细胞成骨分化相关蛋白ALP,RUNX2和OPN蛋白表达促进剂中的用途,其特征在于,所述siRNA为LINC02192的siRNA。
7.根据权利要求6所述的用途,其特征在于,所述LINC02192的序列如SEQ ID NO.1所示;所述siRNA的序列如SEQ ID NO.2和SEQ ID NO.3所示。
8.一种促进牙髓干细胞成骨分化的方法,其特征在于,包括如下步骤:
(1)将原代培养或传代培养获得的牙髓干细胞接种于培养装置中;
(2)将LINC02192的siRNA导入到牙髓干细胞中,加入成骨诱导培养基进行培养。
9.根据权利要求8所述的方法,其特征在于,所述siRNA的序列如SEQ ID NO.2和SEQ IDNO.3所示。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101151371A (zh) * | 2004-12-30 | 2008-03-26 | 高等健康研究院 | 治疗中的逆转录转座子抑制 |
WO2014153548A1 (en) * | 2013-03-21 | 2014-09-25 | The Trustees Of Columbia University In The City Of New York | Compositions and methods for dental tissue regeneration |
CN105561338A (zh) * | 2015-05-14 | 2016-05-11 | 首都医科大学附属北京口腔医院 | Sfrp2在促进牙源性间充质干细胞成骨/成牙分化中的应用 |
CN112239744A (zh) * | 2020-10-20 | 2021-01-19 | 青岛市妇女儿童医院(青岛市妇幼保健院、青岛市残疾儿童医疗康复中心、青岛市新生儿疾病筛查中心) | 一种能够促进牙髓干细胞成骨转化的干扰rna及其应用 |
CN112342217A (zh) * | 2020-11-11 | 2021-02-09 | 杨桂梅 | 一种牙周膜干细胞增殖和成骨分化促进剂 |
CN112410291A (zh) * | 2020-11-24 | 2021-02-26 | 艾冬梅 | 一种牙髓干细胞成牙本质分化诱导剂 |
-
2021
- 2021-11-26 CN CN202111423161.8A patent/CN114107184A/zh active Pending
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101151371A (zh) * | 2004-12-30 | 2008-03-26 | 高等健康研究院 | 治疗中的逆转录转座子抑制 |
WO2014153548A1 (en) * | 2013-03-21 | 2014-09-25 | The Trustees Of Columbia University In The City Of New York | Compositions and methods for dental tissue regeneration |
CN105561338A (zh) * | 2015-05-14 | 2016-05-11 | 首都医科大学附属北京口腔医院 | Sfrp2在促进牙源性间充质干细胞成骨/成牙分化中的应用 |
CN112239744A (zh) * | 2020-10-20 | 2021-01-19 | 青岛市妇女儿童医院(青岛市妇幼保健院、青岛市残疾儿童医疗康复中心、青岛市新生儿疾病筛查中心) | 一种能够促进牙髓干细胞成骨转化的干扰rna及其应用 |
CN112342217A (zh) * | 2020-11-11 | 2021-02-09 | 杨桂梅 | 一种牙周膜干细胞增殖和成骨分化促进剂 |
CN113106097A (zh) * | 2020-11-11 | 2021-07-13 | 青岛思拓新源细胞医学有限公司 | 一种用于牙周膜干细胞成骨分化的促进剂 |
CN112410291A (zh) * | 2020-11-24 | 2021-02-26 | 艾冬梅 | 一种牙髓干细胞成牙本质分化诱导剂 |
Non-Patent Citations (5)
Title |
---|
FU, S.等: "ACCESSION CP034494.1, Eukaryotic synthetic construct chromosome 16", GENBANK, 18 March 2019 (2019-03-18) * |
STRAUSBERG RL等: "ACCESSION NR_110649.1,Homo sapiens long intergenic non-protein coding RNA 2192 (LINC02192), long non-coding RNA", GENBANK, 8 September 2020 (2020-09-08) * |
XINGMEI FENG等: "Repeated lipopolysaccharide stimulation promotes cellular senescence in human dental pulp stem cells (DPSCs)", CELL TISSUE RES, vol. 356, no. 02, 28 March 2014 (2014-03-28), pages 369 - 380 * |
王健雄等: "长链非编码RNA对间充质干细胞成骨分化的影响", 临床内科杂志, vol. 38, no. 09, 15 September 2021 (2021-09-15), pages 645 - 648 * |
王慧君等: "组蛋白去甲基化酶Jmjd3对牙髓干细胞成牙本质向分化的影响研究", 中国实用口腔科杂志, vol. 13, no. 05, 15 May 2020 (2020-05-15), pages 301 - 304 * |
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